Norsyahida A, Rahmah N, Ahmad R M Y
Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 11800 Penang, Malaysia.
Lett Appl Microbiol. 2009 Nov;49(5):544-50. doi: 10.1111/j.1472-765X.2009.02694.x.
To investigate the effects of feeding and induction strategies on the production of BmR1 recombinant antigen.
Fed-batch fermentation was studied with respect to the specific growth rate and mode of induction to assess the growth potential of the bacteria in a bioreactor and to produce high yield of BmR1 recombinant antigen. Cells were grown at a controlled specific growth rate (mu(set)) during pre-induction, followed by constant feeding postinduction. The highest biomass (24.3 g l(-1)) was obtained during fed-batch process operated at mu(set) of 0.15 h(-1), whereby lower mu(set) (0.075 h(-1)) gave the highest protein production (9.82 mg l(-1)). The yield of BmR1 was increased by 1.2-fold upon induction with 1 mmol l(-1) IPTG (isopropyl-beta-d-thiogalactoside) compared to using 5 mmol l(-1) and showed a further 3.5-fold increase when the culture was induced twice at the late log phase.
Combination of feeding at a lower mu(set) and twice induction with 1 mmol l(-1) IPTG yielded the best result of all variables tested, promising an improved method for BmR1 production.
This method can be used to increase the production scale of the BmR1 recombinant antigen to meet the increasing demand for Brugia Rapid(), a commercial diagnostic test for detection of brugian filariasis.
研究补料和诱导策略对BmR1重组抗原生产的影响。
研究了分批补料发酵中特定生长速率和诱导方式，以评估生物反应器中细菌的生长潜力并高产BmR1重组抗原。细胞在诱导前以可控的特定生长速率（μ（设定值））生长，诱导后进行连续补料。在以0.15 h⁻¹的μ（设定值）运行的分批补料过程中获得了最高生物量（24.3 g l⁻¹），而较低的μ（设定值）（0.075 h⁻¹）则产生了最高的蛋白质产量（9.82 mg l⁻¹）。与使用5 mmol l⁻¹相比，用1 mmol l⁻¹异丙基-β-D-硫代半乳糖苷（IPTG）诱导时，BmR1的产量提高了1.2倍，当培养物在对数后期诱导两次时，产量进一步提高了3.5倍。
在所有测试变量中，较低μ（设定值）补料与1 mmol l⁻¹ IPTG两次诱导相结合产生了最佳结果，有望改进BmR1的生产方法。
该方法可用于扩大BmR1重组抗原的生产规模，以满足对Brugia Rapid（一种用于检测布鲁氏丝虫病的商业诊断测试）不断增长的需求。
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